Compositions of Hsp60 Peptides and Viral Antigens for Vaccination and Diagnosis

a technology applied in the field of compositions of hsp60 peptides and viral antigens for vaccination and diagnosis, can solve the problems of rarely avoiding pathogenic response, vaccines generated from complex materials, and inherently having a relatively high probability of inducing competing responses, so as to and increase the immunogenicity of antigens

Inactive Publication Date: 2008-11-13
B G NEGEV TECH & APPL LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0031]The Ec27 peptide is now demonstrated for the first time to increase significantly the immunogenicity of a broad array of antigens, including but not limited to viral antigens, bacterial antigens and mammalian antigens, e.g., viral peptide antigens, bacterial polysaccharides and a...

Problems solved by technology

Despite remarkable achievements in the development of vaccines for certain viral infections (i.e., polio and measles), and the eradication of specific viruses from the human population (e.g., smallpox), viral diseases remain as important medical and public health problems.
Indeed, viruses are responsible for several “emerging” (or re-emerging) diseases (e.g., West Nile encephalitis and Dengue fever), and viral infection is a cause of significant morbidity and mortality worldwide.
The approach suffers from the well known limitations in that it is rarely possible to avoid the pathogenic response because of the complexity of the vaccine which includes not only the antigenic determinant of interest but many related and unrelated deleterious materials, any number of which may, in some or all individuals, induce an undesirable reaction in the host.
These vaccines, generated from complex materials, inherently have a relatively high probability of inducing competing responses even from the antigen of interest.
In immun...

Method used

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  • Compositions of Hsp60 Peptides and Viral Antigens for Vaccination and Diagnosis
  • Compositions of Hsp60 Peptides and Viral Antigens for Vaccination and Diagnosis
  • Compositions of Hsp60 Peptides and Viral Antigens for Vaccination and Diagnosis

Examples

Experimental program
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Effect test

example 1

Natural History of MCMV Dissemination in Spleen and Salivary Gland

[0190]MCMV infection is characterized by different kinetics and viral loads in different organs (Mercer and Spector, 1986). BALB / c mice, 6-8 weeks old, were injected i.p. with 5×104 pfu of MCMV. Mice were sacrificed on days 1, 3, 7, 14 and 28 after infection, and spleens and salivary glands were assayed for infectious virus and MCMV DNA. FIG. 1, shows a typical pattern of MCMV replication in spleen (empty diamonds) and salivary gland (full squares). Virus replication peaked in the spleen on day 3 after infection, and slowly declined thereafter (FIG. 1A). By day 14, no infectious virus could be recovered from this organ. To detect MCMV DNA in infected organs, we used a sensitive PCR using a gB gene primer pair that amplifies a 356 bp segment (Palmon et al., 1996). Viral DNA was detected in the spleen as early as day 1 after infection, peaked on day 3 and by day 14 no DNA could be detected (FIG. 1B).

[0191]In the salivar...

example 2

Immunization with p458-89pep Suppresses MCMV Persistence in the Salivary Gland

[0192]89pep is the H-2Ld-restricted YPHFMPTNL epitope of MCMV-pp 89 (Reddehase et al., 1989). We synthesized chimeric p458-89pep and compared its protective efficacy against MCMV to that induced by 89pep alone or by negative control-89pep. p458 is a MHC class II-restricted peptide derived from murine HSP60 (aa 458-474) and capable of inducing CD4+ T responses in BALB / c mice (Amir-Kroll et al., 2003). The mycobacterial HSP60 431-447 aa peptide (with a val deletion at position 442) did not elicit an immune response to itself or to p458, and thus served as a negative control peptide for immunization; control-89pep.

[0193]To investigate whether immunization with the different peptides would decrease MCMV replication in salivary glands, 3-week old BALB / c mice were immunized twice with IFA alone, 89pep, p458-89pep or control-89pep (FIG. 2). Three-week old, female BALB / c mice were immunized (i.f.p.) with various p...

example 3

IFNγ Secretion by 89pep-Specific T Cells Following Infection and Vaccination

[0196]It is well established that clearance of MCMV during acute infection depends primarily on Th1 IFNγ secretion and protective CTL responses (Mercer and Spector, 1986; Reddehase et al., 1989). We tested whether IFNγ secretion was stimulated by 89pep from spleen (FIG. 3A) and salivary gland (FIG. 3B) cell cultures of MCMV-challenged mice. Cell cultures were prepared on days 1, 3, 7, 14, 21, and 28 after infection, plated for 3 d with or without 89pep, and IFNγ secretion was measured. In the absence of 89pep stimulation, secretion of IFNγ was detected only in spleen cultures from days 1 and 3 after infection. This result probably reflects NK activity in the early stages of infection. When 89pep was added to the cultures, IFNγ secretion in spleen and salivary glands was correlated with the kinetics of viral replication in these organs (FIGS. 1A and 3). It is noteworthy that no significant IL-4 secretion was ...

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Abstract

The present invention provides improved vaccines comprising an isolated viral antigenic peptide and a synthetic peptide derived from a T cell epitope of HSP60. The invention includes mixtures where the peptide serves as an adjuvant as well as conjugates where the peptide is covalently linked to the viral antigen. The known synthetic peptide carrier, p458, provides significantly improved immunogenicity for synthetic viral epitopes and analogs. Ec27 is a novel peptide derived from HSP60 which increases the immunogenicity substantially of the viral antigen both as a mixture or a covalent conjugate. Some of the isolated viral epitopes are novel and are claimed for diagnostic as well as therapeutic or prophylactic uses.

Description

FIELD OF THE INVENTION[0001]The present invention relates to vaccines providing enhanced immunogenicity comprising HSP60 peptides conjugated to or mixed with a viral antigen. The present invention further identifies certain novel epitopes, compositions thereof and methods of using same for vaccination or diagnosis.BACKGROUND OF THE INVENTION[0002]Despite remarkable achievements in the development of vaccines for certain viral infections (i.e., polio and measles), and the eradication of specific viruses from the human population (e.g., smallpox), viral diseases remain as important medical and public health problems. Indeed, viruses are responsible for several “emerging” (or re-emerging) diseases (e.g., West Nile encephalitis and Dengue fever), and viral infection is a cause of significant morbidity and mortality worldwide.[0003]The presence of adequate T-cell help is important for the construction of potent vaccines. Vaccines that induce both helper T cells and CTLs may be more effec...

Claims

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Application Information

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IPC IPC(8): A61K39/245A61K39/12C12N5/10C12N15/31C07K7/00C12N7/00C12Q1/70C12N15/85A61P31/22A61P31/12
CPCA61K2039/6043A61K2039/64G01N33/56983C12N2770/24122A61K47/4833C07K14/005A61K47/646A61P31/12A61P31/22A61P37/04Y02A50/30
Inventor COHEN, IRUN R.RAGER-ZISMAN, BRACHAPORGADOR, ANGELHERKEL, JOHANNES
Owner B G NEGEV TECH & APPL LTD
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